Methods of inhibiting aa9 lytic polysaccharide monooxygenase catalyzed inactivation of enzyme compositions

Abstract

The present invention relates to methods of inhibiting AA9 lytic polysaccharide monooxygenase catalyzed inactivation of an enzyme composition or a component thereof, methods for increasing production of an enzyme composition, and methods for stabilizing an enzyme composition.

1 . A method of inhibiting AA9 lytic polysaccharide monooxygenase catalyzed inactivation of an enzyme composition or a component thereof, said method comprising: adding one or more oxidoreductases selected from the group consisting of a catalase, a laccase, a peroxidase, and a superoxide dismutase to the enzyme composition comprising an AA9 lytic polysaccharide monooxygenase and one or more enzyme components, wherein the one or more added oxidoreductases inhibit AA9 lytic polysaccharide monooxygenase catalyzed inactivation of the one or more enzyme components of the enzyme composition. 
     
     
         2 . The method of  claim 1 , wherein the enzyme composition further comprises one or more components selected from the group consisting of a hydrolase, an isomerase, a ligase, a lyase, an oxidoreductase, or a transferase. 
     
     
         3 . The method of  claim 1 , wherein the enzyme composition further comprises one or more components selected from the group consisting of a cellulase, an AA9 polypeptide, a hemicellulase, a cellulose inducing protein, an esterase, an expansin, a ligninolytic enzyme, a pectinase, a protease, and a swollenin. 
     
     
         4 . The method of  claim 1 , wherein the protein ratio of the added oxidoreductase to the AA9 lytic polysaccharide monooxygenase is in the range of about 1:250 to about 1:10. 
     
     
         5 . The method of  claim 1 , wherein the amount of inhibition of the AA9 lytic polysaccharide monooxygenase catalyzed inactivation is higher in the presence of the one or more added oxidoreductases compared to the absence of the one or more added oxidoreductases. 
     
     
         6 . A method for increasing production of an enzyme composition, said method comprising:
 (a) fermenting a host cell to produce the enzyme composition in the presence of one or more added oxidoreductases selected from the group consisting of a catalase, a laccases, a peroxidase, and a superoxide dismutase, wherein the enzyme composition comprises an AA9 lytic polysaccharide monooxygenase and one or more enzyme components, wherein the one or more added oxidoreductases inhibit the AA9 lytic polysaccharide monooxygenase catalyzed inactivation of the one or more enzyme components of the enzyme composition, and wherein the amount of the enzyme composition produced in the presence of the one or more added oxidoreductases is higher compared to the amount of the enzyme composition produced in the absence of the added one or more oxidoreductases; and   (b) recovering the enzyme composition.   
     
     
         7 . The method of  claim 6 , wherein the host cell comprises an AA9 lytic polysaccharide monooxygenase native to the host cell; an AA9 lytic polysaccharide monooxygenase heterologous to the host cell; or an AA9 lytic polysaccharide monooxygenase native to the host cell and an AA9 lytic polysaccharide monooxygenase heterologous to the host cell. 
     
     
         8 . The method of  claim 6 , wherein the enzyme composition further comprises one or more components selected from the group consisting of a hydrolase, an isomerase, a ligase, a lyase, an oxidoreductase, or a transferase. 
     
     
         9 . The method of  claim 6 , wherein the enzyme composition further comprises one or more components selected from the group consisting of a cellulase, an AA9 polypeptide, a hemicellulase, a cellulose inducing protein, an esterase, an expansin, a ligninolytic enzyme, a pectinase, a protease, and a swollenin. 
     
     
         10 . The method of  claim 6 , wherein the one or more added oxidoreductases are added to the fermentation; the one or more added oxidoreductases are recombinantly produced by the host cell; the one or more added oxidoreductases are recombinantly produced by co-culture of the recombinant cell with a second host cell; the one or more added oxidoreductases are added to the fermentation and recombinantly produced by the host cell; the one or more added oxidoreductases are added to the fermentation and recombinantly produced by co-culture of the recombinant cell with a second host cell; the one or more added oxidoreductases are recombinantly produced by the host cell and recombinantly produced by co-culture of the recombinant cell with a second host cell; or the one or more added oxidoreductases are added to the fermentation, recombinantly produced by the host cell, and recombinantly produced by co-culture of the recombinant cell with a second host cell. 
     
     
         11 . The method of  claim 6 , wherein the protein ratio of the added oxidoreductase to the AA9 lytic polysaccharide monooxygenase is in the range of about 1:250 to about 1:10. 
     
     
         12 . The method of  claim 6 , wherein the inhibition of the AA9 lytic polysaccharide monooxygenase catalyzed inactivation is higher in the presence of the one or more added oxidoreductases compared to the absence of the one or more added oxidoreductases. 
     
     
         13 . A method for stabilizing an enzyme composition, comprising adding one or more oxidoreductases selected from the group consisting of a catalase, a laccases, a peroxidase, and a superoxide dismutase to the enzyme composition, wherein the enzyme composition comprises an AA9 lytic polysaccharide monooxygenase and one or more enzyme components, and wherein the one or more added oxidoreductases inhibit AA9 lytic polysaccharide monooxygenase catalyzed inactivation of the one or more enzyme components of the enzyme composition. 
     
     
         14 . The method of  claim 13 , wherein the enzyme composition further comprises one or more components selected from the group consisting of a hydrolase, an isomerase, a ligase, a lyase, an oxidoreductase, or a transferase. 
     
     
         15 . The method of  claim 13 , wherein the enzyme composition further comprises one or more components selected from the group consisting of a cellulase, an AA9 polypeptide, a hemicellulase, a cellulose inducing protein, an esterase, an expansin, a ligninolytic enzyme, a pectinase, a protease, and a swollenin. 
     
     
         16 . The method of  claim 13 , wherein the protein ratio of the added oxidoreductase to the AA9 lytic polysaccharide monooxygenase is in the range of about 1:250 to about 1:10. 
     
     
         17 . The method of  claim 13 , wherein the amount of inhibition of the AA9 lytic polysaccharide monooxygenase catalyzed inactivation is higher in the presence of the one or more added oxidoreductases compared to the absence of the one or more added oxidoreductases. 
     
     
         18 . (canceled) 
     
     
         19 . (canceled) 
     
     
         20 . (canceled)